17447: Bacillus cellulosilyticus DSM 2522 – results from the first alkaline Bacillus genome

A number of alkaline pretreatments have been proposed for the initial step in the hydrolysis of biomass to 5 carbon and 6 carbon sugars, including alkaline fiber explosion (AFEX), aqueous ammonia, and alkaline peroxide. Cost-effective conversion of the pretreated biomass is limited by the lack of affordable, high specific activity enzymes that function at high pH.  Potential sources of these enzymes are alkaline Bacillus species, including Bacillus cellulosilyticus DSM 2522 (Bce).  The complete genome sequence of Bce was finished by the DOE Joint Genome Institute in July, 2010; yielding a single contig of 4.6 M.  Preliminary analysis indicates that B. cellulosilyticus contains ~40 glycosyl hydrolase genes, only three of which have been described in the literature.  A significant number of the genes contain LPXTG anchor domains, suggesting the  organism uses a combination of soluble and cell-bound enzymes for biomass degradation.  With the full genome sequence available, we will attempt to express and characterize all of the glycosyl hydrolase using a high throughput cloning and expression system developed at Lucigen. The active enzymes will also be sent to partners in the GLBRC to assess their ability to deconstruct plant biomass.